Dendritic cells transfected with DNA constructs encoding CCR9, IL-10, and type II collagen demonstrate induction of immunological tolerance in an arthritis model.

Introduction: Restoring immune tolerance is a promising area of therapy for autoimmune diseases. One method that helps restore immunological tolerance is the approach using tolerogenic dendritic cells (tolDCs). In our study, we analyzed the effectiveness of using dendritic cells transfected with DNA constructs encoding IL-10, type II collagen, and CCR9 to induce immune tolerance in an experimental model of arthritis. Methods: Dendritic cell cultures were obtained from bone marrow cells of Balb/c mice. Dendritic cells (DCs) cultures were transfected with pmaxCCR9, pmaxIL-10, and pmaxCollagen type II by electroporation. The phenotype and functions of DCs were studied using enzyme-linked immunosorbent assay (ELISA) and flow cytometry. Migration of electroporated DCs was assessed in vitro. Induction of antigen-collagen induced arthritis (ACIA) was carried out according to the protocol in Balb/c mice. DCs were then administered to ACIA mice. The development of arthritis was monitored by measuring paw swelling with a caliper at different time points. The immunological changes were assessed by analyzing the content of antibodies to type II collagen using enzyme immunoassay. Additionally, a histological examination of the joint tissue was conducted, followed by data analysis. The results are as follows: DCs were obtained, characterized by reduced expression of CD80, CD86, and H-2Db (MHC class I), increased expression of CCR9, as well as producing IL-10 and having migratory activity to thymus cells. Transfected DCs induced T-regulatory cells (T-reg) and increased the intracellular content of IL-10 and TGF-ß in CD4+T cells in their co-culture, and also suppressed their proliferative activity in response to antigen. The administration of tolDCs transfected with DNA constructs encoding type II collagen, IL-10, and CCR9 to mice with ACIA demonstrated a reduction in paw swelling, a reduction in the level of antibodies to type II collagen, and a regression of histological changes. Conclusion: The study presents an approach by which DCs transfected with DNA constructs encoding epitopes of type II collagen, IL-10 and CCR9 promote the development of antigen-specific tolerance, control inflammation and reduce the severity of experimental arthritis through the studied mechanisms: induction of T-reg, IL-10, TGF-ß.

Immune-checkpoint inhibitor-mediated myocarditis: CTLA4, PD1 and LAG3 in the heart.

Immune-checkpoint inhibitors (ICIs) have revolutionized oncology, with nearly 50% of all patients with cancer eligible for treatment with ICIs. However, patients on ICI therapy are at risk for immune-related toxicities that can affect any organ. Inflammation of the heart muscle, known as myocarditis, resulting from ICI targeting cytotoxic T lymphocyte-associated antigen 4 (CTLA4), programmed cell death protein 1 (PD1) and PD1 ligand 1 (PDL1) is an infrequent but potentially fatal complication. ICI-mediated myocarditis (ICI-myocarditis) is a growing clinical entity given the widespread use of ICIs, its increased clinical recognition and growing use of combination ICI treatment, a well-documented risk factor for ICI-myocarditis. In this Review, we approach ICI-myocarditis from a basic and mechanistic perspective, synthesizing the recent data from both preclinical models and patient samples. We posit that mechanistic understanding of the fundamental biology of immune-checkpoint molecules may yield new insights into disease processes, which will enable improvement in diagnostic and therapeutic approaches. The syndrome of ICI-myocarditis is novel, and our understanding of immune checkpoints in the heart is in its nascency. Yet, investigations into the pathophysiology will inform better patient risk stratification, improved diagnostics and precision-based therapies for patients.

In Vitro Model of Suppression of the Alloantigen Response by Tolerogenic Dendritic Cells Transfected with Personalized DNA Constructs Encoding HLA Epitopes.

BACKGROUND: A search for efficient graft rejection modulation techniques for the promotion of durable engraftment remains to be a matter of close study all over the world. Despite the variety of immunosuppressive drugs, the schemes currently used show a lack of selectivity and have a number of side effects. Here we investigated an approach for the induction of antigen-specific tolerance in a human "stimulator-responder" model in vitro, using dendritic cells (DCs) transfected with designed DNA constructs encoding the stimulator's major histocompatibility complex (MHC) epitopes. METHODS: The object of the study is peripheral blood mononuclear cells (PBMCs) from 10 healthy donors. To induce antigen-specific tolerance, personalized DNA constructs were created for five responder-stimulator pairs, based on the sequences of donors' and recipients' MHCs. DNA sequencing was performed to select epitopes for incorporation into genetic constructs. A mixed lymphocyte culture assay was used (i) to assess the proliferative response in both directions for all possible stimulator-responder pairs (90 reactions) and (ii) to assess the tolerogenic properties of the generated transfected DCs (5 reactions). RESULTS: A significant increase in the amounts of FoxP3+ CD4+CD25+ cells and in IL-10 production was shown in culture of donor mononuclear cells after co-cultivation with the responder's dendritic cells transfected with donor-specific plasmids. The tolerogenic cultures generated using tolerogenic DCs transfected with MHC epitopes had a significantly greater ability to inhibit the proliferation of autologous MNCs in response to an allogeneic MHC stimulus. CONCLUSIONS: The produced DCs transfected with DNA constructs against HLA stimulating epitopes exhibited tolerogenic properties and may be used to develop antigen-specific tolerance. Thus, we proposed a perspective approach to the induction of antigen-specific tolerance, which should subsequently be studied for use in clinical practice.

The cardiac-enriched microprotein mitolamban regulates mitochondrial respiratory complex assembly and function in mice.

Emerging evidence indicates that a subset of RNA molecules annotated as noncoding contain short open reading frames that code for small functional proteins called microproteins, which have largely been overlooked due to their small size. To search for cardiac-expressed microproteins, we used a comparative genomics approach and identified mitolamban (Mtlbn) as a highly conserved 47-amino acid transmembrane protein that is abundantly expressed in the heart. Mtlbn localizes specifically to the inner mitochondrial membrane where it interacts with subunits of complex III of the electron transport chain and with mitochondrial respiratory supercomplexes. Genetic deletion of Mtlbn in mice altered complex III assembly dynamics and reduced complex III activity. Unbiased metabolomic analysis of heart tissue from Mtlbn knockout mice further revealed an altered metabolite profile consistent with deficiencies in complex III activity. Cardiac-specific Mtlbn overexpression in transgenic (TG) mice induced cardiomyopathy with histological, biochemical, and ultrastructural pathologic features that contributed to premature death. Metabolomic analysis and biochemical studies indicated that hearts from Mtlbn TG mice exhibited increased oxidative stress and mitochondrial dysfunction. These findings reveal Mtlbn as a cardiac-expressed inner mitochondrial membrane microprotein that contributes to mitochondrial electron transport chain activity through direct association with complex III and the regulation of its assembly and function.

The sensitivity and specificity of rotational thromboelastometry (ROTEM) to detect coagulopathy during moderate and severe postpartum haemorrhage: a prospective observational study.

INTRODUCTION: Point-of-care viscoelastic haemostatic assays such as rotational thromboelastometry (including ROTEM and TEG) have been used in the management of postpartum haemorrhage (PPH). This study compared results obtained from the automated ROTEM Sigma with laboratory tests of coagulation and platelet count during PPH. METHODS: A prospective observational cohort study recruited women with PPH ≥1000 mL (or clinical concern of bleeding). The Fibtem A5, Extem CT and Pltem (Extem A5 - Fibtem A5) results were compared with laboratory tests of fibrinogen, prothrombin time (PT), activated partial thromboplastin time (APTT) and platelet count. RESULTS: 521 women were recruited, including 274/277 (98.9%) of women with PPH ≥1500 mL. Fibtem A5 results were matched with laboratory fibrinogen in 552/644 (85.7%) samples. The incidence of abnormal laboratory results was low: fibrinogen ≤2 g/L 23/464 (5.0%), PT or APTT >1.5 × midpoint of reference range 4/464 (0.9%), and platelet count <75 × 109/L 11/477 (2.3%). Area-under-the-receiver operator characteristic curve for Fibtem A5 to detect fibrinogen ≤2 g/L was 0.96 (95% Confidence Interval (CI) 0.94 to 0.98, P<0.001), with sensitivity and specificity of Fibtem A5 ≤11 mm to detect fibrinogen ≤2 g/L of 0.76 and 0.96. Prolonged Extem CT results improved after treatment of hypofibrinogenaemia alone. Intervention points for platelet and fresh frozen plasma (FFP) transfusion based on ROTEM Sigma parameters could not be established. CONCLUSION: During PPH (≥1000 mL or cases of clinical concern about bleeding), ROTEM Sigma Fibtem A5 can detect fibrinogen ≤2 g/L and guide targeted fibrinogen replacement. Laboratory results should continue to be used to guide platelet and FFP transfusion.

Cytochrome P450 isoforms 1A1, 1B1 AND 2W1 as targets for therapeutic intervention in head and neck cancer.

Epidemiological studies have shown that head and neck cancer (HNC) is a complex multistage process that in part involves exposure to a combination of carcinogens and the capacity of certain drug-metabolising enzymes including cytochrome P450 (CYP) to detoxify or activate such carcinogens. In this study, CYP1A1, CYP1B1 and CYP2W1 expression in HNC was correlated with potential as target for duocarmycin prodrug activation and selective therapy. In the HNC cell lines, elevated expression was shown at the gene level for CYP1A1 and CYP1B1 whereas CYP2W1 was hardly detected. However, CYP2W1 was expressed in FaDu and Detroit-562 xenografts and in a cohort of human HNC samples. Functional activity was measured in Fadu and Detroit-562 cells using P450-Glo™ assay. Antiproliferative results of duocarmycin prodrugs ICT2700 and ICT2706 revealed FaDu and Detroit-562 as the most sensitive HNC cell lines. Administration of ICT2700 in vivo using a single dose of ICT2700 (150 mg/kg) showed preferential inhibition of small tumour growth (mean size of 60 mm3) in mice bearing FaDu xenografts. Significantly, our findings suggest a potential targeted therapeutic approach to manage HNCs by exploiting intratumoural CYP expression for metabolic activation of duocarmycin-based prodrugs such as ICT2700.

Dendritic Cells Transfected with MHC Antigenic Determinants of CBA Mice Induce Antigen-Specific Tolerance in C57Bl/6 Mice.

BACKGROUND: Nonspecific immunosuppressive therapy for graft rejection and graft-versus-host disease (GVHD) is often accompanied by severe side effects such as opportunistic infections and cancers. Several approaches have been developed to suppress transplantation reactions using tolerogenic cells, including induction of FoxP3+ Tregs with antigen-loaded dendritic cells (DCs) and induction of CD4+IL-10+ cells with interleukin IL-10-producing DCs. Here, we assessed the effectiveness of both approaches in the suppression of graft rejection and GVHD. METHODS: IL-10-producing DCs were generated by the transfection of DCs with DNA constructs encoding mouse IL-10. Antigen-loaded DCs from C57BL/6 mice were generated by transfection with DNA constructs encoding antigenic determinants from the H2 locus of CBA mice which differ from the homologous antigenic determinants of C57BL/6 mice. RESULTS: We found that both IL-10-producing DCs and antigen-loaded immature DCs could suppress graft rejection and GVHD but through distinct nonspecific and antigen-specific mechanisms, respectively. Discussion. We provide data that the novel approach for DCs antigen loading using DNA constructs encoding distinct homologous determinants derived from major histocompatibility complex genes is effective in antigen-specific suppression of transplantation reactions. Such an approach eliminates the necessity of donor material use and may be useful in immunosuppressive therapy side effects prevention.

Impact of traumatic perinatal events on burnout rates among midwives.

BACKGROUND: Little is known about the prevalence of burnout among Irish midwives and how traumatic perinatal events in work contributes to this. AIMS: To establish the prevalence of burnout among midwives in Ireland and whether exposure to traumatic perinatal events in work contributes to this. METHODS: A cross-sectional study utilizing a designed questionnaire was carried out in a tertiary-referral maternity hospital involving all clinical midwives (n = 248). Demographic details and frequency of perinatal events deemed traumatic were recorded. The extent of distress was documented on two visual analogues read in combination to reflect the impact of the distressing events. Burnout severity was assessed using the Copenhagen Burnout Inventory. RESULTS: The response rate was 55% (n = 137). Mean scores for personal, work-related and patient-related burnout were 56.0, 55.9 and 34.3, respectively. Over 90% of respondents experienced exposure to a traumatic event in work in the previous year, with 58% reporting a frequency of monthly or greater. No significant relationship was demonstrated between frequency of trauma and burnout; however, the extent of distress experienced was positively related to burnout in each domain (R2 = 0.18, 0.15 and 0.09, respectively, P < 0.01). A modest negative linear relationship exists between personal and work-related burnout and increasing age (ρ = -0.25 and -0.27, P < 0.01). A significant difference in work-related burnout score was evident between midwives with less experience and more experienced colleagues (P < 0.01). CONCLUSIONS: Burnout is common among midwives. Exposure to discrete traumatic perinatal events experienced by women under their care contributes to this.

Impact of HEXACO Personality Factors on Consumer Video Game Engagement: A Study on eSports.

This article aims to uncover novel insights into personality factors and consumer video game engagement modeling. This research empirically validates the role of specific HEXACO personality factors that foster consumer engagement (CE) in electronic sports (eSports) users. Using a survey-based approach, we incorporated the HEXACO 60 items and consumer video game engagement scales for data collection. Data were collected from eSports users, with 250 valid responses. WarpPLS 6.0 was used for partial least squares-structural equation modeling analyses comprising measurement and structural model assessment. The results showed that the reflective measurement model is reliable and sound, whereas the second-order formative measurement model also meets the criteria of indicator weights and collinearity values variance inflation factor (VIF). The results based on the structural model indicate that openness to experience, extraversion, agreeableness, and conscientiousness positively predict CE in eSports. This article is first among others that conceptualizes and validates the HEXACO personality traits as a reflective formative model using the hierarchical component model approach. The research model carries the explanatory capacity for CE in eSports concerning personality dimensions as indicated by the HEXACO model. It highlights the potential benefits of such research especially to marketers who could potentially employ personality modeling to develop tailored strategies to increase CE in video games.

Work cessation after cancer diagnosis: a population-based study.

BACKGROUND: Long-term work maintenance among cancer survivors is important for patients, their families and society. AIMS: To assess the risk of work cessation among workers at baseline in cancer survivors at 2 and 4 years after diagnosis compared with a matched cancer-free control group. METHODS: Baseline measurements for this historical prospective study were drawn from the Israeli Central Bureau of Statistics 1995 National Census, followed up until 2011. Patients who died before the end of 2011 were excluded from the study. Adjusted odds ratios (ORs) for the study outcome were assessed by binary logistic regression analyses, controlled for age, sex, ethnicity, years of education and socioeconomic position. RESULTS: Cancer was associated with not working at 2 years after diagnosis (adjusted OR = 1.71, 95% confidence interval [CI] 1.59-1.84, P < 0.001), while only mild attenuation was seen at 4 years after diagnosis (adjusted OR = 1.57, 95% CI 1.46-1.68, P < 0.001). Analysis by cancer type revealed that patients diagnosed with central nervous system (adjusted OR = 3.42, 95% CI 2.41-4.86, P < 0.001), renal (adjusted OR = 2.10, 95% CI 1.38-3.16, P < 0.001), breast (adjusted OR = 2.05, 95% CI 1.76-2.38, P < 0.001) and haematologic malignancies (adjusted OR = 2.04, 95% CI 1.59-2.61, P < 0.001) showed the greatest magnitude effect at 2 years. CONCLUSIONS: These results emphasize the need for tailored interventions in order to enhance work maintenance, even among patients who are working at baseline and with very long-term survivors.

Application of a new source model of a panoramic gamma irradiator on dose map formation in an irradiated product.

The computer simulation of radiation processing in an industrial panoramic gamma facility generates a number of complex scientific and technical problems; for example, the product is manually loaded onto turntables and rotates during the entire irradiation and the Monte Carlo MCNP three-dimensional geometry simulation is static. This causes the first problem; for this it is necessary to introduce additional approaches to describe the real irradiation process. It is important to properly represent the irradiation process and the specific conditions of the irradiator to correctly reproduce the measured results and guarantee also through the use of the model developed the most accurate dose map formations in an irradiated product, improving the service quality provided to the customer. The objective of this study is to apply the proposed source model of a panoramic gamma irradiator on dose distributions in containers of different geometric shape of an irradiated product with use of the Monte Carlo code. The influence was studied with use of a new homogenized annular source model. The validation of the MCNPX model of the GammaBeam 127 source was performed by measurements. The validation of the proposed model was based on comparative evaluation of the MCNPX absorbed-dose rates (Gy/h) in the irradiated product between the real source model and the new model. The comparative results for the MCNPX absorbed-dose rate showed that the new source model provides a better description of the irradiation process in a panoramic gamma irradiator. This indicates that the physical and mathematical MCNPX models developed are reliable and correct, and the new source model adequately reproduces the observed dose distributions in the different geometric shapes of the irradiated product.

The DWORF micropeptide enhances contractility and prevents heart failure in a mouse model of dilated cardiomyopathy.

Calcium (Ca2+) dysregulation is a hallmark of heart failure and is characterized by impaired Ca2+ sequestration into the sarcoplasmic reticulum (SR) by the SR-Ca2+-ATPase (SERCA). We recently discovered a micropeptide named DWORF (DWarf Open Reading Frame) that enhances SERCA activity by displacing phospholamban (PLN), a potent SERCA inhibitor. Here we show that DWORF has a higher apparent binding affinity for SERCA than PLN and that DWORF overexpression mitigates the contractile dysfunction associated with PLN overexpression, substantiating its role as a potent activator of SERCA. Additionally, using a well-characterized mouse model of dilated cardiomyopathy (DCM) due to genetic deletion of the muscle-specific LIM domain protein (MLP), we show that DWORF overexpression restores cardiac function and prevents the pathological remodeling and Ca2+ dysregulation classically exhibited by MLP knockout mice. Our results establish DWORF as a potent activator of SERCA within the heart and as an attractive candidate for a heart failure therapeutic.

MOXI Is a Mitochondrial Micropeptide That Enhances Fatty Acid ß-Oxidation.

Micropeptide regulator of ß-oxidation (MOXI) is a conserved muscle-enriched protein encoded by an RNA transcript misannotated as non-coding. MOXI localizes to the inner mitochondrial membrane where it associates with the mitochondrial trifunctional protein, an enzyme complex that plays a critical role in fatty acid ß-oxidation. Isolated heart and skeletal muscle mitochondria from MOXI knockout mice exhibit a diminished ability to metabolize fatty acids, while transgenic MOXI overexpression leads to enhanced ß-oxidation. Additionally, hearts from MOXI knockout mice preferentially oxidize carbohydrates over fatty acids in an isolated perfused heart system compared to wild-type (WT) animals. MOXI knockout mice also exhibit a profound reduction in exercise capacity, highlighting the role of MOXI in metabolic control. The functional characterization of MOXI provides insight into the regulation of mitochondrial metabolism and energy homeostasis and underscores the regulatory potential of additional micropeptides that have yet to be identified.

New source models to represent the irradiation process in panoramic gamma irradiator.

The use of gamma irradiation technologies generates a number of complex scientific and technical problems; for example, the target is manually loaded onto turntables and is rotated during the entire irradiation process and the MCNPX three-dimensional geometry simulation is kept static. For this, it is necessary to introduce additional approaches. In this paper, two new methodologies are proposed for the simulation of irradiation process in panoramic gamma irradiator. The study was performed at the gamma irradiation facility at the Nuclear Technology Development Center of the National Nuclear Energy Commission, Brazil. The source can be reproduced with a homogenized geometry. Validation of MCNPX calculations of gamma doses were performed by thorough comparison with the experimental measurements. The contribution of this proposed source models has opened new lines of research. The results of this study showed that the proposed source models effectively represent the irradiation process.

Genotyping of hepatitis B and C virus Russian isolates for reference serum panel construction.

Approximately 2% and 5% of the world human population is estimated to be infected with HCV and HBV, respectively. Reference panels of HCV and HBV serum samples with defined genotypes and serotypes is necessary for monitoring of the specificity and sensitivity of diagnostic test kits. The aim of this study was to determine genotypes/serotypes of HBV and HCV circulating in Russia in order to construct a panel of reference sera containing these HCV genotypes and HBV serotypes. A total of 343 HBsAg-positive and 207 anti-HCV positive serum samples were collected from patients with HBV and HCV infection from different cities between years 2002 and 2010 in St. Petersburg, Krasnodar, Nizhny Novgorod, Novosibirsk, Barnaul, Gorno-Altaisk, and Khabarovsk. HBV DNA was found in 76.4% of HBsAg positive samples by PCR for the S gene and HCV RNA was found in 71.5, 70.0, and 64.7% of anti-HCV positive samples in the 5'UTR, Core, and NS5B regions, respectively. The prevalence and proportion of HBV genotype/serotype associations were as follows: A/adw2, 2.1%; D/ayw2, 54.0%; D/ayw3, 43.1%; D/adw2, 0.7%. A new combination of genotype D and adw2 serotype was discovered. The distribution of HCV genotypes was the following: 43.6%, b; 3.8%, 2a; and 52.6%, 3a. Russian National reference panels of HBV and HCV lyophilized sera were developed to monitor specificity and sensitivity of approved kits and for the certification of newly developed assays.

Adherence to physical activity guidelines among cancer support group participants.

Physical activity is recommended after cancer diagnosis for physical function, quality of life and survival benefits. This study provided preliminary data on the prevalence of physical activity among adult men and women with cancer in the UK. As part of a national survey of cancer support group participation, questionnaires including items on leisure-time physical activity and demographic information were completed by 748 cancer survivors. Overall, 395 (52.8%) participants reported no weekly moderate or vigorous intensity physical activity, 221 (29.5%) reported some activity but below minimum recommendations and 132 (17.6%) were meeting published guidelines. Gender, health status and socio-economic status were independently associated with meeting guidelines. Among participants in good or fair health who were not meeting guidelines, 59.9% thought that they ought to be more physically active. In conclusion, overall levels of physical activity are low among cancer survivors in the UK. However, the majority of insufficiently active participants showed awareness of the need to increase their activity, and may be receptive to interventions for promoting physical activity in this population.

A UK survey of the impact of cancer on employment.

BACKGROUND: Improvements in cancer detection and treatment and an increase in retirement age mean more people may experience cancer during their working lives. AIMS: To examine the impact of cancer on work activities, sources of advice and support for return-to-work decisions and the role of employers in supporting employees with cancer. METHODS: A cross-sectional survey of a randomly selected sample of people from two cancer registries was conducted in England. Eligible individuals were invited to participate via their general practitioners (April-October 2011) and completed a questionnaire online or by telephone interview. Survey weights were applied before statistical analysis, ensuring responses were representative of cancer survivors in the random sample. RESULTS: A total of 382 people completed the survey, 27% of those invited to participate. Full-time employment fell from 53% prior to diagnosis to 33% after diagnosis, and average working hours reduced from 38 to 32h per week. Only 48% discussed employment issues with their oncology treatment team, and this was associated with more hours worked (36.7 versus 29.4h). Seventy-six per cent of employers were perceived to have been very supportive and 56% receptive to a phased return-to-work. CONCLUSIONS: This is one of the largest UK registry-based surveys on this subject. Following treatment for cancer, there were significant falls in full-time working and hours worked. Just under half the sample discussed employment issues with their treatment team, and these participants worked significantly more hours. This indicates scope for improvement such as encouraging health professionals to raise work-related issues within time-limited consultations.

Informing future research priorities into the psychological and social problems faced by cancer survivors: a rapid review and synthesis of the literature.

PURPOSE: To establish what is known regarding the psychological and social problems faced by adult cancer survivors (people who are living with and beyond a diagnosis of cancer) and identify areas future research should address. METHOD: A rapid search of published literature reviews held in electronic data bases was under taken. Inclusion and exclusion criteria, and removal of duplicated papers, reduced the initial number of papers from 4051 to 38. Twenty-two review papers were excluded on grounds of quality and 16 review papers were selected for appraisal. RESULTS: The psychological and social problems for cancer survivors are identified as depression, anxiety, distress, fear of recurrence, social support/function, relationships and impact on family, and quality of life. A substantial minority of people surviving cancer experience depression, anxiety, and distress or fear associated with recurrence or follow up. There is some indication that social support is positively associated with better outcomes. Quality of life for survivors of cancer appears generally good for most people, but an important minority experience a reduction in quality of life, especially those with more advanced disease and reduced social and economic resources. The majority of research knowledge is based on women with breast cancer. The longer term implications of cancer survival have not been adequately explored. CONCLUSIONS: Focussing well designed research in the identified areas where less is already known about the psychological and social impact of cancer survival is likely to have the greatest impact on the wellbeing of people surviving cancer.